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Quantitative RT-PCR assays for identification and typing of the Equine encephalosis virus

Maan, Sushila; Belaganahalli, Manjunatha N.; Maan, Narender Singh; Potgieter, Abraham C.; Mertens, Peter P. C.

Authors

Sushila Maan

Manjunatha N. Belaganahalli

Narender Singh Maan

Abraham C. Potgieter

Peter P. C. Mertens

Abstract

Equine encephalosis (EE) is an acute, arthropod-borne, noncontagious, febrile disease of equids. The clinical signs of EE are similar to milder forms of African horse sickness (AHS) and the two diseases can be easily confused. The Equine encephalosis virus (EEV) is a distinct virus species within the genus Orbivirus, family Reoviridae, with ten linear segments of dsRNA genome. Seven distinct serotypes of EEV have been recognised on the basis of sequence analyses of Seg-2. The need for differential diagnosis of similar forms of EE and AHS warranted the development of molecular diagnostic methods for specific detection and identification of EEV. We report the development of quantitative real-time RT-PCR assay for detection of any member of the EEV species targeting the highly conserved EEV Seg-9. Similar serotype-specific qRT-PCR assays were designed for each of the seven EEV serotypes targeting genome Seg-2, encoding the serotype determining VP2 protein. These assays were evaluated using different EEV serotypes and other closely related orbiviruses. They were shown to be EEV virus species–specific, or EEV type–specific capable of detecting 1 to 13 copies of viral RNA in clinical samples. The assays failed to detect RNA from closely related orbiviruses, including AHSV and Peruvian horse sickness virus (PHSV) isolates.

Journal Article Type Article
Publication Date Jan 31, 2019
Journal Brazilian Journal of Microbiology
Electronic ISSN 1678-4405
Peer Reviewed Peer Reviewed
Volume 50
Issue 1
Pages 287-296
Institution Citation Maan, S., Belaganahalli, M. N., Maan, N. S., Potgieter, A. C., & Mertens, P. P. C. (2019). Quantitative RT-PCR assays for identification and typing of the Equine encephalosis virus. Brazilian Journal of Microbiology, 50(1), 287-296. doi:10.1007/s42770-018-0034-1
DOI https://doi.org/10.1007/s42770-018-0034-1
Publisher URL https://link.springer.com/article/10.1007/s42770-018-0034-1#enumeration

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